In this application we propose to enhance the newly established Core Microscopy Facilities at the University of New Mexico Health Sciences Center (UNM HSC) through the acquisition of a live cell imaging system. The proposed instrumentation will be integrated with our existing Zeiss LSM 510 and conventional digital imaging equipment. The requested instrumentation includes a new inverted microscope (Axiovert 200) equipped with automated filter wheels and shutter controls, and a fully x, y, z axis motorized stage. Environmental control is provided by micro-observation chambers, objective heaters and perfusion equipment from Bioptechs. For image capture we plan to equip the Axiovert 200 with a digital video camera or transfer our existing LSM51O scanhead to the baseport mount on the Axiovert 200. The LSM51O scanhead is equipped with three lasers (Ar, green HeNe and red HeNe) and is capable of imaging a number of live cell probes and autofluorescent protein variants. For quantification and image analysis we are requesting the Physiology software package to be installed on our Zeiss LSM5 10. Three dimensional imaging software is already installed and with the addition of the Physiology software package the system will perform 4D imaging, allow tracing and quantification of organellar movements, as well as fluorescence resonance energy transfer (FRET) measurements and quantification of fluorescence recovery after photobleaching (FRAP) experiments. The requested digital video camera will be integrated with an existing digital video imaging workstation complete with mage capture and analysis capabilities. The acquisition of the proposed instrumentation will serve seven principal users and several minor users and will offer these independent investigators advanced live cell imaging capabilities important for their NIH-funded biomedical research. Live cell studies will be used to investigate intracellular membrane transport, temporal assembly of membrane microdomains, the regulation of gene expression and host pathogen interactions.